Hypocotyl Extension Research Articles

Functional mechanism study of the allelochemical myrigalone A identifies a group of ultrapotent inhibitors of ethylene biosynthesis in plants

Allelochemicals represent a class of natural products released by plants as root, leaf, and fruit exudates that interfere with the growth and survival of neighboring plants. Understanding how allelochemicals function to regulate plant responses may provide valuable new approaches to better control plant function. One such allelochemical, Myrigalone A (MyA) produced by Myrica gale, inhibits seed germination and seedling growth through an unknown mechanism. Here, we investigate MyA using the tractable model Dictyostelium discoideum and reveal that its activity depends on the conserved homolog of the plant ethylene synthesis protein 1-aminocyclopropane-1-carboxylic acid oxidase (ACO). Furthermore, in silico modeling predicts the direct binding of MyA to ACO within the catalytic pocket. In D. discoideum, ablation of ACO mimics the MyA-dependent developmental delay, which is partially restored by exogenous ethylene, and MyA reduces ethylene production. In Arabidopsis thaliana, MyA treatment delays seed germination, and this effect is rescued by exogenous ethylene. It also mimics the effect of established ACO inhibitors on root and hypocotyl extension, blocks ethylene-dependent root hair production, and reduces ethylene production. Finally, in silico binding analyses identify a range of highly potent ethylene inhibitors that block ethylene-dependent response and reduce ethylene production in Arabidopsis. Thus, we demonstrate a molecular mechanism by which the allelochemical MyA reduces ethylene biosynthesis and identify a range of ultrapotent inhibitors of ethylene-regulated responses.

HSP90 in morphogenesis: taking the heat and keeping the dark.

This article is a Commentary on Zeng et al. (2023), 239: 1253–1265.

Peanut plant growth was altered by monocropping-associated microbial enrichment of rhizosphere microbiome

Repeated planting of the same or closely-related crop species often results in negative soil feedbacks, manifested as reduced plant growth. This is commonly attributed to accumulation of plant pathogenic organisms, but there is increasing evidence that other microbes may contribute as well. Since individual members of the bacterial soil microbiome display distinct preferences for specific root exudates, we hypothesize that enrichment of a small subset of bacterial species in the rhizosphere by monocropping will decrease overall diversity and thus negatively influence the performance of the crop. To test this, we examined soil feedbacks for peanut plants inoculated with bacterial suspensions obtained from monocropped and rotated plots in closed cultivation systems. Partial 16S rRNA gene amplicon sequence analysis revealed significant effects of cropping system on the bacterial composition of peanut rhizospheres. When added to peanut seedling rhizospheres, soil suspensions derived from monocropped plots produced a significant reduction in rhizosphere microbiome species richness (number of OTUs). And, bacterial species including Sphingomonas sp., Herbaspirillum sp., and Arthrobacter sp. were enriched in peanut rhizosphere. However, monocropping-derived soil suspension inoculants showed significant deleterious effects on peanut development compared to rotation-derived inoculants. Further bioassays determined that some enriched bacterial strains that were isolated from the monocropping treatment repressed peanut hypocotyl extension. Our results suggest that bacterial composition assembly in peanut rhizosphere in monocropping system especially that enriches particular deleterious bacterial taxa could lead to clear reductions in plant performance even in the absence of disease or signs of pathogenesis.

Inhibitory Effects of the Lipophilic Extracts and An Isolated Meroditerpene of Brown Alga in Pasture Weeds in the Eastern Amazon Region

ABSTRACT: Two lipophilic extracts and atomaric acid (1), an isolated natural product, were obtained from the marine brown alga Stypopodium zonale (Dictyotaceae) to identify and characterize their potential inhibitory effects on the seed germination, radicle elongation, and hypocotyl development of the weeds Mimosa pudica and Senna obtusifolia. The extracts were prepared with hexane and dichloromethane, and atomaric acid (1) was isolated from hexane extract by way of conventional chromatographic methods. During a 15 days period, germination bioassays were performed at 25 oC with a 12 h photoperiod, whereas radicle elongation and hypocotyl development were assayed at 25 oC with a 24 h photoperiod. After, Petri dishes 9.0 cm in diameter were coated with qualitative filter paper, 25 seeds were placed in a germination chamber, while six pregerminated seeds were placed in the Petri dish for 2-3 days. After 10 days, radicle and hypocotyl extension were measured; and the inhibitory potential of the extracts was assessed at 10 ppm and that of the atomaric acid at 5, 10, 15, and 20 ppm. In both M. pudica and S. obtusifolia, dichloromethane extract achieved the greatest rates of inhibition during seed germination (34% and 22%, respectively), radical germination (38% and 30%, respectively), and hypocotyl development (29% and 22%, respectively). At a concentration of 20 ppm, atomaric acid (1) also demonstrated reduced inhibitory potential, with mean values of 58.67% for M. pudica and 48.67% for S. obtusifolia.

Meta-Analysis of Arabidopsis KANADI1 Direct Target Genes Identifies a Basic Growth-Promoting Module Acting Upstream of Hormonal Signaling Pathways.

An intricate network of antagonistically acting transcription factors mediates the formation of a flat leaf lamina of Arabidopsis (Arabidopsis thaliana) plants. In this context, members of the class III homeodomain leucine zipper (HD-ZIPIII) transcription factor family specify the adaxial domain (future upper side) of the leaf, while antagonistically acting KANADI transcription factors determine the abaxial domain (future lower side). Here, we used a messenger RNA sequencing approach to identify genes regulated by KANADI1 (KAN1) and subsequently performed a meta-analysis combining our data sets with published genome-wide data sets. Our analysis revealed that KAN1 acts upstream of several genes encoding auxin biosynthetic enzymes. When exposed to shade, we found three YUCCA genes, YUC2, YUC5, and YUC8, to be transcriptionally up-regulated, which correlates with an increase in the levels of free auxin. When ectopically expressed, KAN1 is able to transcriptionally repress these three YUC genes and thereby block shade-induced auxin biosynthesis. Consequently, KAN1 is able to strongly suppress shade-avoidance responses. Taken together, we hypothesize that HD-ZIPIII/KAN form the basis of a basic growth-promoting module. Hypocotyl extension in the shade and outgrowth of new leaves both involve auxin synthesis and signaling, which are under the direct control of HD-ZIPIII/KAN.

Arabidopsis cytosolic alpha‐glycan phosphorylase, PHS2, is important during carbohydrate imbalanced conditions

Arabidopsis thaliana has two isoforms of alpha-glycan phosphorylase (EC 2.4.1.1), one residing in the plastid and the other in the cytosol. The cytosolic phosphorylase, PHS2, acts on soluble heteroglycans that constitute a part of the carbohydrate pool in a plant. This study aimed to define a physiological role for PHS2. Under standard growth conditions phs2 knock-out mutants do not show any clear growth phenotype, and we hypothesised that during low-light conditions where carbohydrate imbalance is perturbed, this enzyme is important. Soil-grown phs2 mutant plants developed leaf lesions when placed in very low light. Analysis of soluble heteroglycan (SHG) levels showed that the amount of glucose residues in SHG was higher in the phs2 mutant compared to wild-type plants. Furthermore, a standard senescence assay from soil-grown phs2 mutant plants showed that leaves senesced significantly faster in darkness than the wild-type leaves. We also found decreased hypocotyl extension in in vitro-grown phs2 mutant seedlings when grown for long time in darkness at 6°C. We conclude that PHS2 activity is important in the adult stage during low-light conditions and senescence, as well as during prolonged seedling development when carbohydrate levels are unbalanced.

Why hypocotyl extension mutants need to be characterized at the cell level: a case study of axr3-1.

Since the discovery of auxin, a debate has taken place as to whether the auxin distribution in elongating organs can account for the distinctive cell elongation profiles that have been found. In an attempt to address this important issue, the elongation profiles of cells have been compared in the hypocotyls of wild-type and auxin-hypersensitive axr3-1 Arabidopsis Columbia ecotype seedlings. Clear differences in cell elongation profiles were found in the two types of seedling, whether they were light- or dark-grown. However, it was not possible unambiguously to ascribe the cell elongation profile differences to the proposition that the axr3-1 mutation causes the hypocotyl to be hypersensitive to auxin. The possibility that the abnormal hypocotyl elongation profile of the mutant was a secondary effect, consequent on a more fundamental effect of the axr3-1 mutation, is considered. It is clear from this study that cell elongation and its control needs to be studied at the cell, and not the organ, level. To characterize a mutant as having a short, or long, hypocotyl is inadequate. To determine which factors control the timing and the magnitude of cell elongation requires the demonstration of correlations between the growth rate of cells and their content of regulating substances or their sensitivity to that substance. Studies of the cell elongation profiles of the many hypocotyl length mutants could also be a very effective means of probing the co-ordination of root and shoot elongation.

Antagonistic but complementary actions of phytochromes A and B allow seedling de-etiolation.

Using dichromatic radiation, we show that the actions of phytochromes A and B (phyA and phyB) in Arabidopsis thaliana are antagonistic in mediating red and far-red radiation effects on seedling de-etiolation and yet act in a complementary manner to regulate de-etiolation, irrespective of spectral composition. At low phytochrome photoequilibria inhibition of hypocotyl extension was strong, because of the action of a far-red high-irradiance response mediated by phyA. At high phytochrome photoequilibria inhibition of hypocotyl extension was also strong, because of the action of phyB. At intermediate photoequilibria hypocotyl inhibition was less strong. In their natural environment, this dual action will strongly retard hypocotyl growth and promote cotyledon opening and expansion both in open daylight and under dense vegetation. Overlapping action by phyA and phyB will substantially promote de-etiolation in sparse vegetation. The antagonistic and complementary actions of phyA and phyB, therefore, allow the optimum regulation of seedling growth after emergence from the soil.

Photomorphogenic responses to UV radiation IV: a comparative study of UVB effects on growth and pigment accumulation in etiolated and de-etiolated hypocotyls of wild-type and aurea mutant of tomato Lycopersicon esculentum Mill)

ABSTRACT The question of how de-etiolation of tomato seedling under continuous monochromatic yellow light exerts an influence on UV radiation-induced responses has been studied. Hypocotyl extension and the accumulation of anthocyanins and UV-absorbing compounds was compared in the aurea mutant of tomato and its isogenic wild type. The data of the present paper provide evidence that, during de-etiolation of tomato seedlings, yellow light exerts its effects over seedlings responsiveness to subsequent UV irradiation through several mechanisms: 1) a significant enhancement of shorter UVB wavelength efficiency in both the genotypes; 2) the abolition of UVA -blue light-induced accumulation of UV-absorbing compounds that does not involve pnyA; 3) the disappearance of UVA-blue light-induced hypocotyl growth inhibition that does not involve phyA; 4) higher anthocyanin accumulation rate in response to UV radiation mediated by phyA. Yellow light-induced growth inhibition and accumulation of UV-absorbing compound both mediated by phyA and present only in wild type tomato, appear to be completely separate from the action of UV radiation on the same responses.

Developmental expression and regulation by light of two closely related beta-tubulin genes in Lupinus albus.

We present here the characterization of a Lupinus albus beta-tubulin gene, TubB2, which is closely related to TubB1. Both TubB1 and TubB2 transcripts are present in the embryonic axis of lupin dry seeds. The patterns of developmental expression of TubB1 and TubB2 beta-tubulin genes are strongly correlated. Both genes are expressed at higher levels in hypocotyls of 7-day-old etiolated plants compared to hypocotyls from plants grown under light/dark cycles. When etiolated plants are exposed to continuous white light, differential changes in the steady state levels of the TubB1 and TubB2 mRNAs from hypocotyls are observed. These changes are accompanied by an inhibition of hypocotyl extension.

Free radical accumulation and lipid peroxidation in testas of rapidly aged soybean seeds: a light-promoted process

The role of free radical-induced damage as a cause of loss of vigour in seeds is by no means resolved. In this contribution, the effects of environmental treatments known to reduce viability rapidly were compared with the effects of long-term, low-temperature storage on germination, hypocotyl growth and free radical accumulation and lipid peroxidation in soybean seeds. Accelerated aging was achieved by incubating seeds at 35°C and 1% relative humidity over H2SO4 for up to 69 days in the light and in darkness. In contrast, seeds under long-term storage were maintained at 5°C and 6% moisture content in darkness for up to 6 years. At 35°C there were rapid and significant reductions in rates of seed germination and hypocotyl extension. Loss of viability and declining vigour were associated with increases in lipid peroxidation and free radical build-up but the latter, surprisingly, was largely confined to the testa rather than the cotyledon. Exposure to light greatly enhanced lipid peroxidation and increased organic free radical accumulation in the translucent testas of seeds, but not in the cotyledons. Similar responses to light were recorded in testas detached from seeds. These results show that in soybean the testa is a significant locus of free radical degenerative events induced by high temperature combined with low moisture.

Extension-growth responses and expression of flavonoid biosynthesis genes in the Arabidopsis hy4 mutant.

The hy4 mutant of Arabidopsis thaliana (L.) Heynh. was previously shown to be impaired in the suppression of hypocotyl extension specifically by blue light. We report here that hy4 is altered in a range of blue-light-mediated extension-growth responses in various organs in seedlings and mature plants: it shows greater length of bolted stems, increased petiole extension and increased leaf width and area in blue light compared to the wild type. The hy4 mutant shows decreased cotyledon expansion in both red and blue light compared to the wild type. Anthocyanin formation and the expression of several flavonoid biosynthesis genes is stimulated by blue light in the wild type but to a much lower extent in hy4. The results indicate that the HY4 gene product is concerned with the perception of blue light in a range of extension-growth and gene-expression responses in Arabidopsis.

Heat Tolerance in Phaseolus lunatus

Heat stress on field grown Phaseolus lunatus (lima bean) can have a significant influence on yield. Lima bean crops grown in Delaware typically yield less pounds per acre than the same cultivars grown in California. Part of this effect may be due to extreme heat conditions or fluctuations during Delaware's summers, which can affect blossom and pod set. Our purpose was to analyze the heat tolerance of various cultivars of P. lunatus using quick bioassays and to establish a relationship to yield in greenhouse temperature trials. Two assays were used. The first, a hypocotyl extension assay, consisted of a treatment of germinated seedlings at 25, 35, or 42 for 2 h and observations of hypocotyl extension at 72 and 96 h posttemperature treatments. Three cultivars [`Fordhook' 1072 (heat-sensitive), `Jackson Wonder' (heat-tolerant), and `Early Thorogreen' (heat tolerant)] were analyzed. Initial results indicated that `Jackson Wonder' and `Early Thorogreen' are capable of surviving the 42C heat shock, but `Fordhook 1072' is not. In the second assay, we measured electrical conductivity of a solution containing hypocotyl sections following incubation at various temperatures (R1). Tissue samples then were boiled and conductivity was measured again (R2). The ratios of R1/R2 × 100 were determined as percent injury. Preliminary data suggests that `Jackson Wonder' is more heat-tolerant in this assay than `Fordhook 1072'. Subsequent experiments will analyze the induction of specific heat shock proteins as a function of cultivar-specific heat tolerance.

Coupling of phytochrome B to the control of hypocotyl growth in Arabidopsis.

Etiolated seedlings of the wild-type (WT) and of the phyB-1 mutant of Arabidopsis thaliana (L.) Heynh. were exposed to red-light (R) and far-red light (FR) treatments to characterize the action of phytochrome B on hypocotyl extension growth. A single R or FR pulse had no detectable effects on hypocotyl growth. After 24-h pre-treatment with continuous FR (FRc) a single R, compared to FR pulse inhibited (more than 70%) subsequent hypocotyl growth in the WT but not in the phyB-1 mutant. This effect of FRc was fluence-rate dependent and more efficient than continuous R (Rc) or hourly FR pulses of equal total fluence. Hypocotyl growth inhibition by Rc was larger in WT than phyB-1 seedlings when chlorophyll screening was reduced either by using broad-band Rc (maximum emission 610 nm) or by using narrow-band Rc (658 nm) over short periods (24 h) or with seedlings bleached with Norflurazon. Hourly R or R+FR pulses had similar effects in WT and phyB-1 mutant etiolated seedlings. It is concluded that phytochrome B is not the only photoreceptor of Rc and that the action of phytochrome B is enhanced by a FRc high-irradiance reaction. Complementary experiments with the phyA-201 mutant indicate that this promotion of a phytochrome B-mediated response occurs via co-action with phytochrome A.

Phytochrome, Gibberellins, and Hypocotyl Growth (A Study Using the Cucumber (Cucumis sativus L.) long hypocotyl Mutant)

The possible involvement of gibberellins (GAs) in the regulation of hypocotyl elongation by phytochrome was examined. Under white light the tall long hypocotyl (lh) cucumber (Cucumis sativus L.) mutant, deficient in a type B-like phytochrome, shows an increased "responsiveness" (defined as response capability) to applied GA4 (the main endogenous active GA) compared to the wild type. Supplementing far-red irradiation results in a similar increase in responsiveness in the wild type. Experiments involving application of the precursor GA9 and of an inhibitor of GA4 inactivation suggest that both the GA4 activation and inactivation steps are phytochrome independent. Endogenous GA levels of whole seedlings were analyzed by combined gas chromatography-mass spectrometry using deuterated internal standards. The levels of GA4 (and those of GA34, the inactivated GA4) were lower in the lh mutant under low-irradiance fluorescent light compared with the wild type, similar to wild type under higher irradiance light during the initial hypocotyl extension phase, and higher during the phase of sustained growth, in which extension involved an increase in the number of cells in the upper region. In all cases, growth of the lh mutant was more rapid than that of the wild type. It is proposed that GA4 and phytochrome control cell elongation primarily through separate mechanisms that interact at a step close to the terminal response.

Temperature and water requirements for germination and emergence of lupin

The responses of germination, emergence, and pre-emergent growth of narrow-leafed lupin (Lupinus angustifolius L.) to a range of soil temperatures and matric potentials were determined in 2 replicate experiments with different seedlots and sowing techniques. Although absolute responses to temperature and water availability differed between experiments, the patterns of the responses did not. Optimum temperatures were close to 20�C and base temperatures were 0-3�C. At temperatures >23�C, germination and growth were highly variable and hindered estimation of ceiling (maximum) temperatures; the lowest was about 30�C for germination. Under optimum conditions, germination took 1-1.5 days and emergence from a depth of 4 cm took 4-4.5 days, equating to 75�C. day (base temperature 2�C). The minimum soil matric potential for growth was -1 to -2 MPa, and emergence was more sensitive to low matric potential than was germination. Hypocotyl extension was more sensitive than radicle extension to low matric potential, leading to relatively long roots at the time of emergence.

A Role for the Surrounding Fruit Tissues in Preventing the Germination of Tomato (Lycopersicon esculentum) Seeds

During tomato seed development the endogenous abscisic acid (ABA) concentration peaks at about 50 d after pollination (DAP) and then declines at later stages (60-70 DAP) of maturation. The ABA concentration in the sheath tissue immediately surrounding the seed increases with time of development, whereas that of the locule declines. The water contents of the seed and fruit tissues are similar during early development (20-30 DAP), but decline in the seed tissues between 30 and 40 DAP. The water potential and the osmotic potential of the embryo are lower than that of the locular tissue after 35 DAP also. Seeds removed from the fruit at 30, 35, and 60 DAP and placed ex situ on 35 and 60 DAP sheath and locular tissue are prevented from germinating. Development of 30 DAP seeds is maintained or promoted by the ex situ fruit tissue with which they are in contact. Their germination is inhibited until subsequent transfer to water, and germination is normal, i.e. by radicle protrusion, and viable seedlings are produced, compared with 30 DAP seeds transferred directly to water; more of these seeds germinate, but by hypocotyl extension, and seedling viability is very poor. Isolated seeds at 35 and 60 DAP re-placed in contact with fruit tissues only germinate when transferred to water after 7 d. At 30 DAP, isolated seeds are insensitive to ABA at physiological concentrations in that they germinate as if on water, albeit by hypocotyl extension. At higher concentrations germination occurs by radicle protrusion. Osmoticum prevents germination, but there is some recovery upon subsequent transfer to water. Seeds at 35 DAP are very sensitive to ABA and exhibit little or no germination, even upon transfer to water. The response of the isolated seeds to osmoticum more closely approximates that to incubation on the ex situ fruit tissues than does their response to ABA. This is also the case for isolated 60 DAP seeds, whose germination is not prevented by ABA, but only by the osmoticum; these seeds are inhibited when in contact with ex situ fruit tissues also. It is proposed that the osmotic environment within the tissues of the tomato fruit plays a greater role than endogenous ABA in preventing precocious germination of the developing seeds.

Role of the Cotyledons in the Maintenance of Hypocotyl Growth in Helianthus annuus L.

Summary The effect of cotyledon excision on hypocotyl growth was investigated in sunflower seedlings (Helianthus annuus L.) that were grown in continuous white light. Removal of both cotyledons resulted in an inhibition of hypocotyl elongation and cessation of primary leaf development. Cell osmotic pressure, the amount of soluble sugars and wall plasticity declined in the hypocotyls without cotyledons and the accumulation of cell wall material stopped. In contrast, the excision of one cotyledon did not affect hypocotyl extension and the development of the primary leaves. Cell osmotic pressure was unaffected in the growing region of the stem, in spite of the fact that the accumulation of soluble sugars was reduced by 50 %. After the removal of one cotyledon a transient inhibition of the accumulation of cell wall material occurred, but no effect on cell wall extensibility was detectable. It is concluded that cell elongation and maintenance of wall plasticity can occur without net increase in cell wall material.

RESPONSES OF LIGHT‐GROWN WILD‐TYPE and LONG‐HYPOCOTYL MUTANT CUCUMBER SEEDLINGS TO NATURAL and SIMULATED SHADE LIGHT*

AbstractThe photomorphogenic control of hypocotyl extension growth was characterized in wild type (WT) and long hypocotyl (Ih) mutant seedlings of cucumber (Cucumis sativus L.) grown under natural radiation in outdoor and glasshouse experiments. Hypocotyl extension growth of WT plants was promoted by supplementing sunlight with far‐red light during the photoperiod, by reducing the amount of blue light reaching either the whole shoot or the hypocotyl, and by reducing the amount of UV reaching the whole shoot.The Ih seedlings only responded to a reduction in UV‐B levels. Both WT and Ih seedlings showed phototropic responses to the direction of blue light. Increasing degrees of vegetational shade promoted hypocotyl growth of WT plants. The Ih mutant showed no hypocotyl growth promotion by natural shade in glasshouse experiments (no UV‐B, low water demand) and a reduced response (10‐23% of the WT response, according to pretreatment conditions) in outdoor experiments (UV‐B, high water demand).

Photoresponses of transgenic tobacco plants expressing an oat phytochrome gene

The physiological responses of transgenic tobacco (Nicotiana tabacum L.) plants that express high levels of an introduced oat (Avena sativa L.) phytochrome (phyA) gene to various light treatments are compared with those of wild-type (WT) plants. Seeds, etiolated seedlings, and light-grown plants from a homozygous transgenic tobacco line (9A4) constructed by Keller et al. (EMBO J, 8, 1005-1012, 1989) were treated with red (R), far-red (FR), or white light (WL) with or without supplemental FR light, revealing major perturbations of the normal photobiological responses. White light stimulated germination of both WT and transgenic seed, but addition of FR to the WL treatment suppressed germination. In the WT, all fluence rates tested inhibited germination, but in the transgenics, reduction effluence rate partially relieved germination from the FR-mediated inhibition. It is suggested that the higher absolute levels of the FR-absorbing form of phytochrome (Pfr) in the irradiated transgenics, compared to the WT, may be responsible for the reduced FR-mediated inhibition of germination in the former. Hypocotyl extension of dark-grown seedlings of both WT and transgenic lines was inhibited by continuous R or FR irradiation, typical of the high-irradiance response (HIR). After 2 d of de-etiolation in WL, the WT seedlings had lost the FR-mediated inhibition of hypocotyl extension, whereas it was retained in the transgenics. The FR-mediated inhibition of hypocotyl extension in the transgenic seedlings after de-etiolation may reflect the persistence of an, FR-HIR response mediated by the overexpressed oat PhyA phytochrome. Light-grown WT seedlings exhibited typical shade-avoidance responses when treated with WL supplemented with high levels of FR radiation. Internode and petiole extension rates were markedly increased, and the chlorophyll a∶b ratio decreased, in the low-R: FR treatment. The transgenics, however, showed no increases in extension growth under low-R: FR treatments, and at low fluence rates both internode and petiole extension rates were significantly decreased by low R ∶ FR. Interpretation of these data is difficult. The depression of the chlorophyll a∶b ratio by low R ∶ FR was identical in WT and transgenic plants, indicating that not all shade-avoidance responses of light-grown plants were disrupted by the over-expression of the introduced oat phyA gene. The results are discussed in relation to the proposal that different members of the phytochrome family may have different physiological roles.